淫羊藿苷激活Nrf2信号通路保护D-半乳糖诱导损伤小鼠睾丸支持细胞15P-1作用初探

Protective effect of icariin on D-galactose induced 15P-1 cells injury via Nrf2 signaling pathway

从Nrf2信号通路探讨淫羊藿苷(icariin,ICA)对D-半乳糖(D-galactose,D-Gal)诱导的小鼠睾丸支持细胞株15P-1细胞损伤的保护作用。将15P-1细胞分为对照组(control)、150 mM D-Gal处理组、D-Gal+ICA(0.5μM)组和D-Gal+ICA(1.0μM)组。RT-PCR法和Western blot法检测15P-1细胞分泌的相关因子GDNF、BMP4和SCF mRNA和蛋白的表达水平。Western blot法检测15P-1细胞紧密连接相关蛋白Occludin和Claudin-1、Nrf2信号通路相关蛋白Nrf2、HO-1和NQO-1的蛋白表达水平。免疫荧光法检测Nrf2表达及定位。与对照组相比,D-Gal处理组15P-1细胞分泌的相关因子GDNF、BMP4和SCF的mRNA和蛋白表达水平均显著下降,而给予ICA后均显著升高。此外,与对照组相比,D-Gal处理组15P-1细胞Occludin、Claudin-1、Nrf2、HO-1和NQO-1的蛋白表达水平均显著下降,而ICA可显著上调上述相关蛋白的表达。免疫荧光结果进一步显示,ICA可上调15P-1细胞核内Nrf2蛋白表达。ICA可减轻D-Gal诱导的15P-1细胞损伤,并改善其功能,其机制可能与上调Nrf2信号通路相关蛋白有关。

To study the protective effect of icariin(ICA)on D-galactose(D-Gal)induced 15 P-1 Sertoli cells injury via the Nrf2 signaling pathway.15 P-1 cells were divided into control group,150 mM D-Gal group,D-Gal+ICA(0.5μM)group and D-Gal+ICA(1.0μM)group.The mRNA and protein expression levels of GDNF,BMP4 and SCF in 15 P-1 cells were detected by RT-PCR and Western blot,respectively.The expression levels of tight junction related proteins Occludin and Claudin-1,Nrf2 signaling pathway proteins including Nrf2,HO-1 and NQO-1 in 15 P-1 cells were detected by Western blot.The expression and localization of Nrf2 were detected by immunofluorescence.The mRNA and protein expression levels of GDNF,BMP4 and SCF in 15 P-1 cells in D-Gal stimulated group were significantly decreased compared with the control group.On the contrary,ICA significantly increased the mRNA and protein expression levels of GDNF,BMP4 and SCF.In addition,the protein expression levels of Occludin,Claudin-1,Nrf2,HO-1,and NQO-1 in 15 P-1 cells in D-Gal group were significantly decreased compared with the control group,while icariin significantly up-regulated the above related protein expression.The results of immunofluorescence further showed that ICA up-regulated the expression of Nrf2 in the nucleus of 15 P-1 cells.ICA can alleviate the injury of 15 P-1 Sertoli cells induced by D-Gal and improve its function,and its mechanism may be related to upregulation of Nrf2 signaling pathway proteins.